CN102617705B

CN102617705B - Macrocyclic compound for suppressing replication of hepatitis c viruses

Info

Publication number: CN102617705B
Application number: CN201210034872.0A
Authority: CN
Inventors: 李本; 陈力; 翟培彬; 江涛
Original assignee: SHANGHAI VIRO MEDICAL Co Ltd
Current assignee: Ginkgo Pharma Co ltd
Priority date: 2012-02-16
Filing date: 2012-02-16
Publication date: 2014-12-31
Anticipated expiration: 2032-02-16
Also published as: EP2816054A1; KR20140134273A; EP2816054A4; WO2013120371A1; JP2015508761A; US9321809B2; JP6105632B2; CA2864488A1; CN102617705A; US20150031603A1; AU2012370125A1; IN2014MN01637A; IL233899A0; KR102004381B1; CN104169293A

Abstract

The invention relates to a compound which is inhibitor for replication of one or multiple types of hepatitis c viruses. In addition, the invention discloses a medicinal component and preparation containing the compound and application of the inhibitor for replication of the hepatitis c viruses. The compound can be used individually or used with other compounds to treat symptoms caused by infection of the hepatitis viruses.

Description

Suppress the macrocyclic compounds that hepatitis C virus copies

Technical field

The present invention relates to compound, prepare the method for compound, the pharmaceutical composition of compound and medicament, and this compound be used for the treatment of, prevent, the disease of diagnosing one or more relevant to hepatitis C virus or symptom.

Background technology

It is the major cause that hepatic diseases is brought out in the whole world that hepatitis C virus (HCV) infects, estimate according to the World Health Organization (WHO), there is the chronic hepatitis C infection person of 1.7 hundred million to 2 hundred million in the current whole world, accounts for 3% of population in the world, and annual newly-increased third hepatopath 3,000,000 ~ 4,000,000 people.Although acute third liver clinical manifestation is comparatively light, easily develop into chronic, the patient of about 50-80% can develop into chronic hepatitis even liver cirrhosis and liver cancer, it is reported, to infect after the third liver 20 years, liver cirrhosis occurs as 10-15%.Current third liver mortality ratio comes the tenth in all diseases in the whole world, and in China, the third liver mortality ratio comes the 5th.

At present, the standard treatments of the third liver is that Peg-Intron (PEG-IFN) share with ribavirin.But from continued viral response rate (SVR), current this standard treatments effect is not very desirable, and the clinical cure rate for 1a/1b type patient is about 50%.And the administration time of current this therapy is long, third hepatopath of such as HCV I type, need continuous use 48 weeks, also often there is serious untoward reaction simultaneously, as with spiritual aspect problem, occur flu-like symptoms and produce haematics toxicity, thus cause the successful curative ratio of existing therapy to be less than 10%, therefore, develop a kind of new mechanism, more the HCV inhibitor of high-efficiency low-toxicity seem particularly important.

HCV genome is a kind of single stranded RNA (+) of flaviviridae, the common 3009-3030 of an about 9600 base-pairs code amino acid whose polypeptide.This polypeptide is cut into by proteolytic enzyme the albumen that 10 have difference in functionality, comprising core protein---Core, envelope glycoprotein---E1, E2, non-structural protein---NS2, NS3 (has serine protease, helicase activity), NS4A, NS4B, NS5A, NS5B (there is RdRP activity), and the albumen of 1 Unknown Function---p7 (finding that it may be a kind of ionic channel recently) is in protein maturation process, Core, E1, cutting between E2 and p7 relies on intracellular signal peptidase to complete, NS2 and NS3 then relies on the cysteine protease activity of self to realize autocatalysis fracture, cutting between all the other albumen is completed by the NS3 after maturation.(Michael P.Manns et al.，Nature Reviews Drug Discovery，6，991-1001(2007))。

Most of NS 3 proteinase inhibitor is enzyme substrates competitor, and its action target spot is substrate binding site.NS3 proteinase inhibitor is large lopps endogenous peptide analogue (product peptides) the earliest, although up to now, this compounds does not go through to go on the market, and has at present and is much in clinical investigation phase.The second NS3 proteinase inhibitor is the plan peptide of linear structure, and its mechanism of action is introduce an electrophilic α carbonylamino group on broken site, makes itself and Serine form a reversible covalent linkage.

The NS3 proteinase inhibitor gone on the market at present has telaprevir and boceprevir, but all the interior life of body is active low for these two kinds of medicines, medicine is general for character, thus cause treating the large shortcoming of consumption, therefore exploitation is efficient and have good medicine becomes exploitation HCV protease inhibitor at present Main way for the proteinase inhibitor of character.

Summary of the invention

One object of the present invention is the treatment providing a kind of compound of suppression hepatitis c virus infection completely newly for HCV infection, provides a kind of brand-new treatment means, for treatment hepatitis c virus infection provides new selection.

Another object of the present invention is to provide a kind of medicinal compositions, it is characterized in that, described composition contains described formula (I) compound or its pharmacy acceptable salt of pharmaceutically acceptable carrier and pharmaceutical effective amount.

Another object of the present invention is to provide a kind of described formula (I) compound or its pharmacy acceptable salt preparing the application in the medicine suppressing hepatitis C virus to copy.

4th object of the present invention is to provide the application in the medicine of preparation control hepatitis c virus infection of a kind of described formula (I) compound or its pharmacy acceptable salt.

In a first aspect of the present invention, provide a kind of formula I or its pharmacy acceptable salt, solvate or prodrug:

Wherein:

R ₁be selected from-CO ₂r _a,-CONR _bsO ₂r _c,-CONR _dsO ₂nR _er _f, or tetrazyl;

R ₂be selected from C1-C6 alkyl, C2-C6 thiazolinyl, or C3-C8 cycloalkyl; Described group can be replaced by 1-3 halogenic substituent;

R ₃be selected from C1-C8 alkyl, C3-C8 cycloalkyl, the alkyl of the C1-C8 replaced by C3-C8 cycloalkyl, or the alkyl of the C1-C8 replaced by aryl and heterocyclic radical; Described group can be replaced by the substituting group that 1 to 3 is selected from lower group: halogen, hydroxyl, nitro, cyano group, trifluoromethyl, trifluoromethoxy ,-NR _ar _b,-SO ₂r _c,-SOR _c,-SR _c,-SO ₂nR _dr _e,-CONR _fr _g,-COOR _h,-NR _icOR _j,-NR _ksO ₂r _l, C1-C6 alkyl, C1-C6 alkoxyl group or C3-C7 cycloalkyl;

R ₄be selected from H, C1-C6 alkyl ,-SO ₂r _c,-SO ₂nR _dr _e,-CONR _fr _g,-COOR _h, or-COR _i;

N is 1 or 2;

P is 0,1 or 2;

M is-O-,-S-or-NH-;

L is selected from C1-C6 methylene radical, C2-C6 thiazolinyl, or C2-C6 alkynyl;

W is selected from halogen, hydroxyl, nitro, cyano group, trifluoromethyl, trifluoromethoxy ,-NR _ar _b,-SO ₂r _c,-SOR _c,-SR _c,-SO ₂nR _dr _e,-CONR _fr _g,-COOR _h,-NR _icOR _j,-NR _ksO ₂r _l, C1-C6 alkyl ,-O-C1-C6 alkyl, C3-C7 cycloalkyl, aryl, or heteroaryl;

Z is selected from C1-C6 methylene radical ,-O-,-O-C1-C5 methylene radical ,-C (O) O-, C1-C5 methylene radical-C (O) O-,-C (O) NR _ar _b-, or C1-C5 methylene radical-C (O) NR _ar _b-

Ring A is selected from 8-14 unit and merges dicyclo or three ring carbon systems, and can by 1-4 N, O, S hybrid atom MCM-41;

In above-mentioned, R _a, R _b, R _c, R _d, R _e, R _f, R _g, R _h, R _i, R _j, R _k, R _lindependently selected from hydrogen, C1-C6 alkyl, C3-C7 cycloalkyl, C5-C10 aryl or heteroaryl, C1-C6 methylene radical C5-C10 aryl or heteroaryl.

In one embodiment, its structure of compound provided by the invention is such as formula shown in (IIa):

In another embodiment, its structure of compound provided by the invention is such as formula shown in (IIb):

In a second aspect of the present invention, provide formula (I) compound or its pharmacy acceptable salt, solvate or prodrug that the invention described above that described in a kind of medicinal compositions, composition contains pharmaceutically acceptable carrier and pharmaceutical effective amount provides.

Preferably, described pharmaceutical composition comprises the second therapeutical agent further, and described second therapeutical agent is selected from HCV antiviral agent, immunomodulator and anti-infective medicament; Described HCV antiviral agent is selected from HCV protease inhibitor and HCV NS5B AG14361.

In one embodiment, the form of described pharmaceutical composition is aqueous dispersant, liquid, Gel mile, syrup, elixir, medicine slurry, suspension, aerosol, control-released agent, quick-dissolving agent, effervescent, freeze-dried, tablet, powder, pill, sugar-coat are complete, capsule, delayed release agent, extend releasing agent, pulse control-released agent, multiparticulates agent or releasing agent immediately.

In a third aspect of the present invention, provide a kind of formula provided by the invention (I) compound as above or its pharmacy acceptable salt, solvate or prodrug are preparing the purposes in medicine, and this medicine is used for prevention or treatment HCV infection in the main body needed.

Accordingly, this invention exploits efficient and there is the HCV-Ab IgG virus compound of good medicine for character.

Embodiment

Contriver, through extensive and deep research, has found the compound that a class effectively can suppress hepatitis C virus (HCV) and copies.

Compound

In one aspect of the invention, a kind of type I compound or its pharmacy acceptable salt is provided, solvate or prodrug:

Wherein,

N is 1 or 2;

P is 0,1 or 2;

M is-O-,-S-or-NH-;

In one aspect of the invention, R ₁be selected from-CONR _bsO ₂r _cand/or R ₄be selected from-COOR _h.

In one aspect of the invention, formula (I) compound have formula (II a) structure:

Wherein, R ₁, R ₂, R ₃, R ₄, n, p, M, L, W as above illustrate.

In one aspect of the invention, formula (I) compound have formula (II b) structure:

Wherein, R ₁, R ₂, R ₃, R ₄, n, p, M, L, W as above illustrate.

Group for different variable described above any is combined in this and expects.

Formula (II a) compound include but not limited to the explanation in table 1.

Table 1

Formula (I) compound includes but not limited to the explanation in table 2.

Table 2

The synthesis of compound

Formula described above (I) compound can use the method combined in the synthetic technology of standard or known technology and literary composition to synthesize.In addition, solvent as mentioned herein, temperature and other reaction conditionss can change.

Starting material for the synthesis of formula (I) compound can by synthesizing or obtaining from commercial source, as, but be not limited to Aldrich Chemical Co. (Milwaukee, Wis.) or Sigma Chemical Co. (St.Louis, Mo.).Compound as herein described and other allied compounds with different substituents can use known technology and raw material to synthesize, and comprise and are found in March, Advanced Organic Chemistry 4th Ed., (Wiley 1992); Carey and Sundberg, Advanced Organic Chemistry 4th Ed., Vols.A and B (Plenum 2000,2001), Green and Wuts, Protective Groups in Organic Synthesis 3rd Ed., the method in (Wiley 1999).General method prepared by compound changes by using suitable reagent and introduce not isoplastic condition in this molecular formula provided.

Compound II per a synthesizes according to flow process I

Flow process I: the synthesis of the large ring plate section A9 in the left side

First, aromatic amine compound A1 and the condensation of 3-L-Hydroxyproline methyl ester compd A 2 generate carbamate A3.Then dipeptides A5 is obtained with amino acid A4 condensation after removing the Boc protecting group on nitrogen.Dipeptides A5 removes nitrogen-protecting group subsequently, then obtains the diene precursors A7 of Guan Huan with Segment A 6 condensation.Diene A7 closes ring by olefin metathesis reaction and obtains compound A-28, namely obtains the large ring plate section A9 on the left side after A8 hydrolysis.

Compound II per b synthesizes according to flow process II

Flow process II: the synthesis of the large ring plate section B7 in the left side

Synthesis and the A9 of the left side large ring plate section B7 are similar, and the 1-chloro-7-bromine isoquinoline namely first replaced from starting raw material is given repeated exhortations B1, obtains intermediate B 2 with 3-aminoproline methyl esters generation substitution reaction.B2 generates B3 by Stille coupling subsequently.There is a series of reaction similar with preparation A9 in B3, obtains fragment B7 after closing cyclizing hydrolysis again.

Flow process III: the synthesis of the right fragment C7

Reference (J.Org.Chem.2005,70,5869-5879), obtains imines C2 by after phenyl aldehyde and glycine ethyl ester mixed dehydration, and adding alkali subsequently and obtaining trans with Isosorbide-5-Nitrae-dibromo 2,3-butene reaction is main cyclopropane derivative C3.Then acid adding removes benzyl and obtains raceme C4 with Boc acid anhydrides protection amino.Optically pure (1R, 2S) ethyl amino acid ester C5 is obtained after splitting with esterase.Obtain C6 with various sulphonamide condensation after LiOH saponification, after removing Boc protection with trifluoromethyl subsequently, obtain the right fragment C7.

Flow process IV: the synthesis of target compound IIa

The large ring plate on the left side is broken A9 and the right amine fragment condensation after can obtain Compound II per a.The reactions such as further hydrogenation or generation addition can obtain the analogue of IIa further.

Flow process V: the synthesis of target compound IIb

The large ring plate on the left side is broken B6 and the right amine fragment condensation after can obtain Compound II per b.The reactions such as further hydrogenation or generation addition can obtain the analogue of IIb further.

The synthesis of formula (I) compound is summarized in an embodiment.

The further form of compound

Should be clear that, some formula (I) compound can present tautomerism.Formula (I) compound can exist with the form of non-solvation, also can exist with the form of solvation.Even, can there is heteromorphism in some formula of the present invention (I) compound.

The pharmacy acceptable salt be applicable to of formula (I) compound can be the acid salt of formula (I) compound, can include, but is not limited to the salt formed with following mineral acid: example hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and the salt formed with organic acid, organic acid then refers to acetic acid, oxalic acid, succinic acid, tartrate, methylsulfonic acid and toxilic acid; Can be the salt with enough acid formula (I) compound, as an alkali metal salt or alkaline earth salt (calcium salt, magnesium salts or ammonium salt etc.).The pharmacy acceptable salt that the another kind of formula (I) compound is applicable to can be the salt formed in human or animal body after giving construction (I) compound.This compound also can with the form of " prodrug " of ester, carbamate or other routines (when administration in this form, can change into active part) in vivo.

Term

If without illustrating in addition, for the present patent application, comprising the term in specification sheets and claims, being defined as follows.Must be noted that, in the specification and the appended claims, if Wen Zhongwu clear instruction in addition, singulative " " comprises plural references.If without illustrating in addition, use the ordinary method of mass spectrum, nuclear-magnetism, HPLC, protein chemistry, biological chemistry, recombinant DNA technology and pharmacology.In this application, if without illustrating in addition, use "or" or " with " refer to "and/or".

" formula (I) compound " refers to that structural formula is (I), and (II a), (II compound b).

" alkyl " refer to straight or branched saturated, aliphatic hydrocarbon group containing 1-8 carbon atom (preferably 1-6 carbon atom); C1-n alkyl then represents the saturated aliphatic radical of 1-n carbon atom, comprise straight chain and branched group (such as " C1-20 alkyl ", refer to that this group is alkyl, and the carbochain amount of carbon atom of alkyl is between 1 ~ 20, namely containing 1 carbon atom, 2 carbon atoms, 3 carbon atoms etc., until comprise the alkyl of 20 carbon atoms.And the restriction of this 1-20 does not comprise the carbonatoms of the replacement on alkyl, as replaced " alkyl " in alkylamino, when being not particularly limited its carbonatoms, only refer to that the carbonatoms of the moieties wherein indicated is 1-20, and the substituent carbonatoms do not comprised on alkyl and other the substituent carbonatomss on amino.The statement of " C1-8 alkyl " is adopted then to represent the alkyl containing 1 ~ 8 carbon atom in this alkyl.) " thiazolinyl " comprise straight chain containing at least one carbon-carbon double bond and 2-8 carbon atom (preferably 2-6 carbon atom) and branched hydrocarbyl; " alkynyl " comprises straight chain containing at least one carbon carbon triple bond and 2-8 carbon atom (preferably 2-6 carbon atom) and branched hydrocarbyl.Haloalkyl, represent the alkyl that halogen atom replaces, this replacement comprises monosubstituted and polysubstituted, and wherein the concept of alkyl is described above.C1 ~ 8 haloalkyl, refers to that the carbonatoms of the alkyl in haloalkyl is 1 ~ 8.Haloalkyl refers to the group that on alkyl, H atom is replaced by halogen atom; As perfluoroalkyl refers to H on alkyl all by group that F replaces.

Term " aralkyl " refers to-alkyl-aryl-group, and wherein alkyl and aryl are as defined herein.

Term " aryl " as used at this refers to aromatic systems, can be monocycle or originally condense or many aromatic rings of linking together, thus make the ring condensing at least partially or connect form the virtue system of conjugation.Aromatic yl group includes, but are not limited to: phenyl, naphthyl, tetralyl.Aryl can be optionally substituted, as can by 1-4 to be selected from the group of lower group the aryl that replaces or heterocycle: halogen, CN, OH, NO ₂, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxyl group, aryloxy, the alkoxyl group of replacement, alkyl-carbonyl, alkyl carboxyl, alkylamino or arylthio.Preferably, substituting group is halogen, C1-C4 alkyl.Term " cycloalkyl " refers to monocycle or polycyclic aliphatic hydrocarbon, non-aromatic free radical, and wherein each atom (as skeletal atom) of Cheng Huan is carbon atom.Cycloalkyl can be saturated or part is unsaturated.Cycloalkyl can be connected with aromatic ring, and tie point is on the carbon atom of non-aromatic ring carbon atom.Cycloalkyl comprises from 3 to 10 ring member nitrogen atoms.In some is concrete, cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl and ring octyl group.Cycloalkyl can be substituted or not replace.

Term " halogen " or " halogenide " refer to fluorine, chlorine, bromine or iodine.

Term " heterocyclic radical " refers to containing the heteroatomic cyclic group by 3 to 8 annular atomses such as N, O, S, and in this group, heteroatoms only containing atom N, also can contain O or S atom.Wherein heteroatomic number can be one, also can be multiple.This heterocycle can be saturated class cycloalkane structure, also can be undersaturated aromatic ring class formation.More specifically, this term nitrogen heterocycle includes but not limited to pyrryl, Pyrrolidine base, piperidyl, piperazinyl, morpholinyl, piperazinyl, pyrimidyl, imidazolyl etc.Heterocycle also can comprise any many rings, and wherein arbitrary above-mentioned heterocycle can condense in aromatic ring.

The chemical bond that term " key " or " singly-bound " refer between two atoms or between two segments (when the atom connected by key is considered to macrostructure a part of).On the one hand, when group as herein described is a key, lack with reference to group, allow to determine to form a key between group remaining.

Term " ring " comprises any ring texture.Term " unit " means the quantity representing the skeletal atom forming ring.Like this, e.g., cyclohexyl, pyridyl, pyranyl, thiapyran base is six-ring, cyclopentyl, pyrryl, and furyl and thienyl are five-rings.

Term " segment " refers to concrete part or the functional group of molecule.Chemistry segment is considered to be included in or attached chemical entities in the molecule usually.

Term " optionally replace " or " replacements " refer to reference group can replace by one or more extra group, extra group is individually and be independently selected from, alkyl, cycloalkyl; aryl, heteroaryl, heterolipid cyclic hydrocarbon, hydroxyl; alkoxyl group, alkylthio, arylthio, alkane sulfoxide group; virtue sulfoxide group, alkane sulfuryl, fragrant sulfuryl, cyano group; halogen, carbonyl, thiocarbonyl, nitro; alkylhalide group, fluoroalkyl and amino, comprise monosubstituted and disubstituted amino group and protected derivative thereof.Illustrate, optional replacement can be halogenide,-CN ,-NO2 or LsRs, wherein each Ls is independently selected from a key,-O-,-C (=O)-,-C (=O) O-,-S-,-S (=O)-,-S (=O) 2-,-NH-,-NHC (=O)-,-C (=O) NH-, S (=O) 2NH-,-NHS (=O) 2 ,-OC (=O) NH-,-NHC (=O) O-, or-(C1-C6 alkyl); Each Rs is selected from hydrogen, alkyl, fluoroalkyl, assorted alkyl, cycloalkyl, aryl, heteroaryl or Heterocyclylalkyl.The protecting group that can form above substituent protection derivative can with reference to Greene and Wuts.On the one hand, optional substituting group is selected from halogen, CF3, OH, CN, NO2, SO3H, SO2NH2, SO2Me, NH2, COOH, CONH2, alkoxyl group ,-N (CH3) 2 and alkyl.

In some specific embodiment, described compound has one or more three-dimensional structure center, and each center independently exists with R or S type.Compound comprises all diastereomeric bodily forms as mentioned herein, the mapping bodily form, and difference is to the structure bodily form and their suitable mixture.Steric isomer obtains by the such as method of chiral chromatographic column to Enantiomer separation.

Method as herein described and molecular formula comprise use N-oxide compound (if suitable), the pharmacy acceptable salt of crystallized form (being also referred to as polymorphic) or formula (I) structural compounds, and the active metabolite of these compounds with identical activity.In some cases, compound may exist as tautomer.All tautomers comprise within the scope of compound as mentioned herein.In certain specific embodiment, described compound with solvation form exist, pharmaceutically acceptable solvent as water, ethanol etc.In other specific embodiment, described compound exists with nonsolvated forms.

Shortenings:

DMF=N, dinethylformamide

NMR=nuclear magnetic resonance spectroscopy(NMR spectroscopy)

LDA=diisopropylamino lithium

THF=tetrahydrofuran (THF)

PE=sherwood oil

EA=ethyl acetate

MS=mass spectroscopy

DCM=methylene dichloride

MeOH=methyl alcohol

DMSO=methyl-sulphoxide

MCPBA=metachloroperbenzoic acid

HOBt=1-hydroxybenzotriazole

DBU=1,5-diazabicylo [5.4.0] 11-5-alkene

NADPH=Triphosphopyridine nucleotide, reduced

ACN=acetonitrile

Specific pharmacy and medical terminology

Term " acceptable ", as used herein, refer to that a prescription component or the health of activeconstituents to general treatment target do not have undue harmful effect.

Term used herein " HCV infection " refers to that HCV virus arrives new organism through blood or other circulation way, and enters organic host cell, starts self-replacation and propagation.

As the term is employed herein " HCV suppression " refer to that compound can reduce vitro culture with replication that the is HCV virus of human infection and Marmot fur.The copy number being embodied in the HCV genetic material RNA in host cell or in blood reduces.

Term " Combined Preparation " or its similar terms, as used herein, refer to by several selected medicine to patient's medication, with identical or different administering mode at identical or different time administration.

Term " experimenter " or " patient " comprise Mammals and nonmammalian.Mammals includes but not limited to, mammals: people, non-human primates are as orangutan, ape and monkey class; Agricultural animal is as ox, horse, goat, sheep, pig; Domestic animal is as rabbit, dog; Laboratory animal comprises rodents, as rat, mouse and cavy etc.Animal includes but not limited to non-mammalian, bird, fish etc.In a preference, selected Mammals is people.

Term " treatment ", " therapeutic process " or " therapy " are as used herein, comprise mitigation, suppress or improve symptom or the situation of disease; Suppress the generation of complication; Improve or prevent potential metabolic syndrome; Suppress the generation of disease or symptom, as controlled the development of disease or situation; Palliate a disease or symptom; Disease or symptom are gone down; Alleviate the complication caused by disease or symptom, or prevent or treat the sign caused by disease or symptom.

As used herein, a certain compound or pharmaceutical compositions pharmaceutical compositions, after administration, a certain disease, symptom or situation can be made to improve, and espespecially its severity improves, and delayed onset slows down disease progression, or reduces the state of an illness time length.No matter fix administration or interim administration, continue medication or interrupted continuous administration, can owing to or the situation relevant with administration.

Therepic use and route of administration

The present invention also comprises pharmaceutical composition and methods for the treatment of, and it comprises to the formula I of administration significant quantity.Compound of the present invention can be used for treatment: HCV infection.Preferably, described Mammals is people.

When compound is used for such use; they can with one or more pharmaceutically acceptable carrier or mixed with excipients; as solvent, thinner etc.; and can with following form oral administration: tablet, capsule, dispersible powder, particle or suspension (containing 0.05-5% suspension agent according to appointment), syrup (containing 10-50% sugar according to appointment) and elixir (containing 20-50% ethanol of having an appointment), or carry out parenteral routes with sterile injectable solution or form of suspension (containing 0.05-5% suspension agent of having an appointment in isotonic medium).Such as, these pharmaceutical preparations containing the about 25-90% mixed with carrier, can be about the activeconstituents of 5%-60% (weight) usually.

The effective dose of activeconstituents used can change with the severity of the pattern of compound used, administration and disease to be treated.But, usually when compound of the present invention gives with the dosage of about 0.5-500mg/kg the weight of animals every day, gratifying effect can be obtained, preferably give with the dosage that 2-4 time is separated every day, or with sustained release forms administration.For most of large mammal, the total dose of every day is about 1-100mg, is preferably about 2-80mg.Be applicable to the dosage form taken orally, comprise the active compound with the intimately mixed about 0.5-500mg of solid-state or liquid pharmaceutically acceptable carrier.This dosage adjustable is replied to provide optimal treatment.Such as, by an urgent demand for the treatment of situation, the dosage that several times separate can be given every day, or dosage is reduced pari passu.

These active compounds are by oral and intravenously, intramuscular or the administration such as subcutaneous.Solid-state carrier comprises: starch, lactose, Si Liaodengji dicalcium phosphate feed grade, Microcrystalline Cellulose, sucrose and white bole, and liquid carrier comprises: sterilized water, polyoxyethylene glycol, nonionic surface active agent and edible oil (as Semen Maydis oil, peanut oil and sesame oil), as long as be applicable to the characteristic of activeconstituents and required specific administration mode.In pharmaceutical compositions, normally used adjuvant also can advantageously be included, and such as seasonings, pigment, sanitas and antioxidant are as vitamin-E, vitamins C, BHT and BHA.

From the position being easy to preparation and administration, preferred pharmaceutical composition is solid-state composition, especially the capsule of tablet and solid-filling or liquid filling.The oral administration of compound is preferred.

These active compounds also can parenteral or intraperitoneal administration.Also solution or the suspension of these active compounds (as free alkali or pharmacy acceptable salt) can be prepared in the water being suitably mixed with tensio-active agent (as hydroxypropylcellulose).Also can prepare dispersion liquid in glycerine, liquid, polyoxyethylene glycol and the mixture in oil thereof.Under conventional storage and working conditions, contain sanitas in these preparations to prevent microorganism growth.

The medicament forms being adapted to inject comprises: aseptic aqueous solution or dispersion liquid and aseptic powder (for extemporaneous preparation of sterile injection solution or dispersion liquid).In all situations, these forms must be aseptic and must be that fluid is to be easy to syringe displacement fluids.Must be stable under conditions of manufacture and storage, and must can prevent the pollution effect of microorganism (as bacterium and fungi).Carrier can be solvent or dispersion medium, wherein containing, for example water, alcohol (as glycerine, propylene glycol and liquid polyethylene glycol), their suitable mixture and vegetables oil.

The present inventor finds after deliberation, macrocyclic compounds Late Cambrian of the present invention, and it can suppress HCV virus efficiently.When being exposed to by hepatitis C virus in the compound of effective concentration, hepatitis C virus can effectively be killed.So compound of the present invention may be used for, for the preparation of the medicine for the treatment of HCV infection, certainly, can comprising pharmaceutically acceptable carrier in medicine.

The all features (comprising any described claim, summary and figure) be described in this manual, and/or the institute related in any method or process in steps, all likely exist with any one combination, unless some feature or step are mutually repel in same combination.

Relative to scheme of the prior art, advantage of the present invention is:

The present invention have found a kind of compound of prevention and therapy hepatitis c virus infection, and making us unexpected is that it has and very strong kills hepatitis C virus effect.When being cultivated together with infecting the described cell of hepatitis C virus by this compound, can find that it can disturb at least some composition of hepatitis C virus in cell.

The present invention have found a kind of compound of brand-new molecular structure, and it is active that it has strong anti-hepatitis C virus, and have outstanding medicine for character.

These embodiments only supply the object of illustrations, are not limited to the scope of the claim that this provides.

General introduction.1H-NMR spectrum uses Bruker-400 nuclear magnetic resonance analyser, and the unit of chemical shift is 1,000,000/, interior mark is tetramethylsilane.Coupling constant (J) is close to 0.1Hz.The shortenings used illustrates as follows: s, singlet; D, doublet; T, triplet; Q, quartet; Qu, quintet; M, multiplet; Br, spectrum.Mass spectrum uses Waters 2795 with the substance quadrupole mass spectrometer of electron spray ionisation (ESI).Silica gel is used to carry out column chromatography.

Embodiment 1 Compound II per a-1

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

Synthesize according to the method for flow process I, III, IV.

Intermediate A 3:(2S, 4R)-4-(4-vinyl isoindole-2-ketonic oxygen base) tetramethyleneimine-1-t-butyl formate 2-methyl-formiate

According to flow process I, raw material A 2 (2.21g, 9.03mmol) is dissolved in DMF (20mL), at 0 DEG C, adds CDI (1.47g, 9.03mmol) in batches.Stirring at room temperature is after 18 hours, slowly drip A1 (prepared by reference literature, J.Me d.Chem.2010,53,2443-2463) (1.4g, 9.03mmol) DMF solution, drip finish be heated to 60 DEG C after stir 2 hours.After being cooled to room temperature, adding frozen water and 5%KHSO4 (35mL) solution successively, be then extracted with ethyl acetate three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains A3 (2.3g, 61%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ7.41-7.38(m，1H)，7.30-7.27(m，1H)，7.18-7.12(m，1H)，6.70(dd，J＝17.6，10.8Hz，1H)，5.73(dd，J＝17.6，4.0Hz，1H)，5.39(d，J＝10.8Hz，1H)，5.34(brs，1H)，4.79-4.67(m，4H)，4.52-4.37(m，1H)，3.78-3.76(m，5H)，2.54-2.43(m，1H)，2.28-2.20(m，1H)，1.47-1.44(m，9H).

Intermediate A 5:(3R, 5S)-1-((S)-2-(t-butoxycarbonyl amino)-3,3-dimethylbutanoyl)-5-(methoxycarbonyl) pyrrolidyl-3-position 4-vinyl isoindole-2-manthanoate

According to flow process I, raw material A 3 (2.2g, 5.28mmol) be dissolved in 4N HCl/dioxane (30mL), stirring at room temperature is after 18 hours, and reaction terminates, and concentrates to obtain de-Boc crude product (1.8g, 100%).Without purifying, directly drop into next step reaction.

By N-Boc-L-Terleu (2.4g, 10.0mmol), de-Boc crude product (3.5g, 10.0mmol), HATU (5.7g, 14.9mmol) and DiPEA (1.9g, 14.9mmol) mixing is dissolved in (40mL) in methylene dichloride.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains brown oil A5 (2.9g, 50%) after column chromatography.

ESI-MS m/z 530.2(M+H) ⁺。

Intermediate A 6:(S)-3-(allyl group oxygen base)-2-(t-butoxycarbonyl amino) propionic acid

Under room temperature, in methylene dichloride (10mL) solution of N-Boc-L-serine methylester (200mg, 0.91mmol) and allyl bromide 98 (552mg, 4.56mmol), add silver suboxide (654mg, 2.74mmol).Lucifuge stirs 3 hours, filters, concentrated, obtains pale yellow oil A6-1 (149mg, 63.1%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ：7.19(d，J＝8.0Hz，1H)，5.89-5.80(m，1H)，5.26(dd，J＝17.6，1.6Hz，1H)，5.16(d，J＝11.6Hz，1H)，4.25(dd，J＝13.6，5.6Hz，1H)，3.94(d，J＝6.8Hz，2H)，3.63(s，3H)，3.61-3.59(m，2H)，1.39(s，9H).

Intermediate A 6-1 (120mg, 0.46mmol) is added in THF/H2O (5mL/2.5mL) solution of LiOH (53mg, 1.28mmol), stirs 1 hour.PH 1N HCl is adjusted to 4, and water layer is separated, and adds extraction into ethyl acetate twice.Saturated common salt is washed, Na ₂sO ₄drying, obtains pale yellow oil A6 (110mg, 97.3%) after filtering and concentrating.Without purifying, be directly used in condensation.

Intermediate A 7:(3R; 5S)-1-((S)-2-((S)-3-(allyloxy)-2-(t-butoxycarbonyl amino)-3,3-dimethylbutanoyl)-5-(methoxycarbonyl) pyrrolidyl-3-position 4-vinyl isoindole-2-manthanoate

According to flow process I, by raw material A 5 (2.9g, 5.5mmol) be dissolved in methylene dichloride (50mL), then drip in trifluoroacetic acid (10mL, 20%), stirring at room temperature is after 3 hours, reaction terminates, add water and methylene dichloride after concentrated, and regulate pH to 12 with 2N NaOH, extract three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating must take off Boc crude product (2.0g, 85.1%).Without purifying, directly drop into next step reaction.

By amino acid A6 (1.1g, 4.7mmol), de-Boc crude product (2.0g, 4.7mmol), HATU (2.3g, 6.1mmol) and DiPEA (0.9g, 7.0mmol) mixing is dissolved in (30mL) in methylene dichloride.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains pale yellow oil A7 (1.25g, 40.3%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ7.41-7.38(m，1H)，7.24-7.14(m，2H)，6.71-6.62(m，0.5H)，5.91-5.81(m，1H)，5.72-5.65(m，0.5H)，5.42-5.36(m，2H)，5.30-5.15(m，3H)，4.86-4.61(m，5H)，4.55-4.51(m，1H)，4.22-4.13(m，2H)，4.00-3.88(m，4H)，3.45-3.38(m，1H)，2.54-2.48(m，1H)，2.27-2.20(m，1H)，1.47-1.43(m，9H)，1.00(s，9H).

Intermediate A 8:N-[(1R, 12E, 17S, 20S, 23S)-20-tertiary butyl-23-(methanoyl methyl)-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

According to flow process I; under nitrogen protection; raw material A 7 (31mg, 0.047mmol) is dissolved in methylene dichloride (20mL), then adds Zhan catalyst (3.5mg; 0.005mmol); after stirred overnight at room temperature, reaction terminates, and adds 0.1mL DMSO cancellation reaction; after concentrated, column chromatography obtains brown oil A8 (10mg, 34.4%).

¹H NMR(400MHz，CDCl ₃)δ7.31-7.25(m，2H)，7.20-7.15(m，1H)，6.83(d，J＝16.0Hz，1H)，6.08(d，J＝16.0Hz，1H)，5.58(d，J＝7.6Hz，1H)，5.29(brs，1H)，4.90-4.65(m，5H)，4.48-4.32(m，4H)，4.17-4.09(m，2H)，3.80-3.76(m，4H)，3.43-3.38(m，1H)，2.78-2.72(m，1H)，2.20-2.14(m，1H)，1.49(s，9H)，1.10(s，9H).

Intermediate A 9:N-[(1R, 12E, 17S, 20S, 23S)-20-tertiary butyl-23-(formyl hydroxyl)-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

According to flow process I, intermediate A 8 (66mg, 0.11mmol) is added in THE/H2O (5mL/2.5mL) solution of LiOH (14mg, 0.33mmol), stirs 1 hour.PH 1N HCl is adjusted to 4, and water layer is separated, and adds extraction into ethyl acetate twice.Saturated common salt is washed, Na ₂sO ₄drying, obtains pale yellow oil A6 (56mg, 86.8%) after filtering and concentrating.Without purifying, be directly used in condensation.

Intermediate C2:(E)-2-(benzyl imines) ethyl acetate

According to flow process III; under nitrogen protection, glycine ethyl ester (65.0g, 0.64mol) and triethylamine (100mL, 0.71mol) are dissolved in toluene (500mL); add phenyl aldehyde (50.0g, 0.47mol) stirring and refluxing 4 hours afterwards.Concentrated, add 200mL and 200mL ethyl acetate, extract three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating obtains brown oil intermediate C2 (80.0g, 88.9%).Without purifying, directly drop into next step reaction.

Intermediate C3:(E)-1-(benzyl imines)-2-vinylcyclopropane ethyl formate

According to flow process III; by intermediate C2 (40.0g under nitrogen protection; 0.20mol) He 1; 4-dibromo 2-butylene (44.0g; 0.20mol) be dissolved in toluene (100mL); slowly drip the toluene solution (100mL) of potassium tert.-butoxide (32.0g, 0.40mol) under stirring, continue stirring 1 hour.Add water and ethyl acetate (20mLx3), extract three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating obtains intermediate C3.Without purifying, directly drop into next step reaction.

Intermediate C4:1-(t-butoxycarbonyl amino)-2-vinylcyclopropane ethyl formate

According to flow process III, intermediate obtained in the previous step is dissolved in ethyl acetate (100mL), stirs the lower toluene solution (100mL) dripping 1N HCl (200mL), continue stirring 2 hours.Stratification, organic layers with water extracting twice.Wash by ethyl acetate after merging aqueous phase, then water layer NaOH regulates pH to 12.Add MTBE, after extracting twice, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating obtains deprotection crude intermediate.This intermediate is dissolved in MTBE again, adds (Boc) ₂o (68.0g, 0.30mol) and triethylamine (50mL, 0.30mol) stirring at room temperature 2 hours afterwards.Add water and ethyl acetate (20mLx3), extract three times.Organic phase 1N HCl washes, and saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains pale yellow oil C4 (26.0g, 49.1%) after column chromatography.

¹H NMR(400MHz，DMSO-d6)δ5.80-5.51(m，1H)，5.30-5.09(m，2H)，4.19-4.13(m，2H)，2.16(q，J＝10.8Hz，1H)，1.80-1.75(m，1H)，1.50-1.45(m，1H)，1.44(s，9H)，1.27(t，J＝6.8Hz，3H)，.

Intermediate C5:(1R, 2S)-1-(t-butoxycarbonyl amino)-2-vinylcyclopropane ethyl formate

According to flow process III, Alcalase 2.4L (100mL) is dissolved in (500ml) in the damping fluid of 40 DEG C, and regulates pH to 8.0 with 50%NaOH.At such a temperature, DMSO (100mL) solution of intermediate C4 (26.0g, 0.11mol) is slowly dripped.Drip and finish, continue stirring 72 hours.By pH regulator to 8.5, product is extracted with ethyl acetate twice.Saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, pale yellow oil C5 (13.0g, 49.1%, 100%ee) after column chromatography.

Intermediate C6:(1R, 2S)-1-(cyclopropanesulfonamide)-2-vinylcyclopropane carbamate

According to flow process III, intermediate C5 (6.0g, 23.5mmol) is added in THF/MeOH/H2O (20mL/20mL/10mL) solution of LiOH (3.0g, 73.5mmol), stirs 1 hour.PH 1N HCl is adjusted to 4, and water layer is separated, and adds extraction into ethyl acetate twice.Saturated common salt is washed, Na ₂sO ₄drying, obtains pale yellow oil after filtering and concentrating.Without purifying, be directly used in condensation.

This intermediate (5.6g, 24.2mmol) and CDI (5.2g, 31.5mmol) are dissolved in THF (20mL).Reflux and be cooled to room temperature after 1 hour, then add methylene dichloride (30mL) solution of cyclopropane base sulphonamide (3.8g, 31.5mmol).Add DBU (5.2mg, 34.0mmol) subsequently, room temperature for overnight.Extraction into ethyl acetate is added twice after concentrated.Saturated common salt is washed, Na ₂sO ₄drying, obtains white solid C6 (2.8g, 36.4%) after filtering and concentrating column chromatography.

¹H NMR(400MHz，CDCl ₃)δ9.52(s，1H)，5.66-5.57(m，1H)，5.32(d，J＝13.2Hz，1H)，5.17(dd，J＝10.4，0.8Hz，1H)，2.94-2.87(m，1H)，2.17(q，J＝8.4Hz，1H)，1.92-1.89(m，1H)，1.48(s，9H)，1.45-1.39(m，1H)，1.32-1.25(m，2H)，1.13-1.00(m，2H).

Intermediate C7:(1R, 2S)-1-amino-2-vinylcyclopropane carbamate

According to flow process III, intermediate C6 (1.0g, 3.0mmol) is dissolved in methylene dichloride (10mL), drips trifluoroacetic acid (2mL, 20%) and stir 2 hours afterwards.Brown oil C7 (1.2g, 100%) is obtained after concentrated.Without purifying, be directly used in condensation.

Compound II per a-1

According to flow process IV, by left side acid fragment A9 (56mg, 0.10mmol), amine fragment C7 (24mg, 0.10mmol), HATU (58mg, 0.15mmol) be dissolved in (10mL) in methylene dichloride with DiPEA (27mg, 0.20mmol) mixing.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains white powder IIa-1 (17mg, 22.7%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ9.91(brs，1H)，7.42-7.38(m，1H)，7.19-7.07(m，2H)，6.75(d，J＝20.8Hz，1H)，6.02(d，J＝20.8Hz，1H)，5.61-5.55(m，2H)，5.23-5.05(m，3H)，4.80-4.64(m，4H)，4.39-4.26(m，5H)，4.10-3.95(m，2H)，3.66-3.64(m，4H)，3.40-3.32(m，1H)，2.82-2.72(m，1H)，2.50-2.40(m，1H)，2.35-2.20(m，1H)，2.00-1.80(m，4H)，1.40(s，9H)，1.35-1.30(m，2H)，1.26-1.15(m，2H)，0.98(s，9H)；ESI-MS m/z 849.00(M+Na) ⁺。

Embodiment 2 Compound II per a-2

(1R, 12E, 17S, 20S, the 23S)-17-amino-20-tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy-s-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides

By Compound II per a-1 (10mg, 0.012mmol) be dissolved in methylene dichloride (5mL), then trifluoroacetic acid (1mL is dripped, 20%) in, stirring at room temperature is after 2 hours, and reaction terminates, and adds water and methylene dichloride after concentrated, and regulate pH to 12 with 2N NaOH, extract three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, white solid IIa-2 (2.3mg, 26.1%) after column chromatography.

ESI-MS m/z 727.00(M+H) ⁺。

Embodiment 3 Compound II per a-3

N-[(1R, 12E, 17R, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

According to the method in embodiment 1, Compound II per a-3 can be prepared from N-Boc-D-serine methylester.

¹H NMR(400MHz，CDCl ₃)δ9.97(brs，1H)，7.28-7.08(m，3H)，6.55(d，J＝17.6Hz，1H)，5.90-5.65(m，3H)，5.52(s，1H)，5.28-5.14(m，2H)，4.82-4.60(m，6H)，4.47-4.26(m，6H)，3.94-3.79(m，2H)，3.42-3.32(m，1H)，2.92-2.82(m，1H)，2.50-2.36(m，2H)，2.11-1.96(m，3H)，1.52(s，9H)，1.35-1.30(m，3H)，1.06(s，9H)，1.06-1.02(m，2H)，；ESI-MS m/z 849.00(M+Na) ⁺。

Embodiment 4 Compound II per a-4

(1R, 12E, 17R, 20S, the 23S)-17-amino-20-tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy-s-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides

According to the method in embodiment 2, after Compound II per a-3 is removed Boc, purifying can obtain Compound II per a-4.

ESI-MS m/z 727.00(M+H) ⁺。

Embodiment 5 Compound II per a-5

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy--17-C-pyrazine-2-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17,23-diamide bases

Compound II per a-2 and 2-pyrazine carboxylic acid condensation purifying can be prepared Compound II per a-5.

¹H NMR(400MHz，CDCl ₃)δ9.46(s，1H)，8.80(d，J＝2.8Hz，1H)，8.72(d，J＝8.0Hz 1H)，8.56(s，1H)，7.75(d，J＝8.0Hz 1H)，7.59(brs，1H)，7.26-7.23(m，2H)，7.19(d，J＝6.8Hz，1H)，6.75(d，J＝17.2Hz，1H)，6.05-6.00(m，1H)，5.78-5.69(m，1H)，5.38(s，1H)，5.28-5.14(m，2H)，5.05-5.01(m，1H)，4.91-4.87(m，1H)，4.80-4.76(m，3H)，4.59-4.51(m，2H)，4.36-4.33(m，2H)，4.19-4.07(m，2H)，3.85-3.81(m，1H)，3.68-3.65(m，1H)，2.90-2.86(m，1H)，2.65-2.60(m，1H)，2.40-2.33(m，1H)，2.19-2.00(m，1H)，1.96-1.92(m，1H)，1.47-1.41(m，1H)，1.10-1.02(m，11H)；ESI-MS m/z 855.00(M+Na) ⁺。

Embodiment 6 Compound II per a-6

(1R, 12E, 17R, 20S, 23S)-20-the tertiary butyl-23-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy--17-C-pyrazine-2-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17,23-diamide bases

Compound II per a-4 and 2-pyrazine carboxylic acid condensation purifying can be prepared Compound II per a-6

¹H NMR(400MHz，CDCl ₃)δ10.0(s，1H)，9.43(s，1H)，8.78(s，1H)，8.73(d，J＝7.8Hz，1H)，8.55(s，1H)，7.63(d，J＝7.8Hz，1H)，7.45(s，1H)，7.26-7.16(m，2H)，6.75(d，J＝16.0Hz，1H)，6.06-6.01(m，1H)，5.78-5.72(m，1H)，5.35(s，1H)，5.28-5.15(m，2H)，4.92-4.74(m，6H)，4.51-4.13(m，7H)，3.85-3.77(m，2H)，3.68-3.62(m，2H)，2.90-2.86(m，1H)，2.65-2.58(m，1H)，2.38-2.33(m，1H)，2.18-1.94(m，4H)，1.47-1.41(m，2H)，1.02(s，9H)；ESI-MS m/z 855.00(M+Na) ⁺。

Embodiment 7 Compound II per a-7

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-17-acetylaminohydroxyphenylarsonic acid 3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides

According to the method in embodiment 5, Compound II per a-2 and Acetyl Chloride 98Min. condensation purifying can be prepared Compound II per a-7.

¹H NMR(400MHz，CDCl ₃)δ10.0(brs，1H)，7.75-7.70(m，2H)，7.24-7.22(m，2H)，7.16(d，J＝6.8Hz，1H)，6.69(s，1H)，6.75(d，J＝16.8Hz，1H)，5.98(d，J＝16.4Hz，1H)，5.68-5.60(m，1H)，5.34(s，1H)，5.30-5.11(m，2H)，4.86-4.74(m，5H)，4.55-4.49(m，2H)，4.32-4.25(m，2H)，3.82-3.80(m，1H)，3.46-3.40(m，1H)，2.85-2.82(m，1H)，2.59-2.50(m，1H)，2.35-2.31(m，1H)，2.17(s，3H)，1.91-1.80(m，2H)，1.40-1.33(m，4H)，1.05(s，9H)；ESI-MS m/z 791.00(M+Na) ⁺。

Embodiment 8 Compound II per a-8

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-17-methanesulfonamido-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides

According to the method in embodiment 5, Compound II per a-2 and methylsulfonyl chloride condensation purifying can be prepared Compound II per a-8.

¹H NMR(400MHz，CDCl ₃)δ10.0(brs，1H)，7.45(d，J＝7.6Hz，1H)，7.34(s，1H)，7.24-7.20(m，2H)，7.15(d，J＝6.8Hz，1H)，6.59(d，J＝16.4Hz，1H)，5.91(d，J＝16.4Hz，1H)，5.78-5.60(m，1H)，5.52(d，J＝8.8Hz，1H)，5.35(s，1H)，5.23-5.12(m，2H)，4.77-4.73(m，4H)，4.52(d，J＝8.0Hz，1H)，4.40-4.19(m，5H)，4.11-4.03(m，2H)，3.81-3.77(m，1H)，3.54-3.40(m，1H)，2.89-2.82(m，1H)，2.59-2.53(m，1H)，2.35-2.28(m，1H)，2.17(s，3H)，2.08-2.00(m，1H)，1.96-1.90(m，1H)，1.40-1.33(m，1H)，1.07(s，9H)，1.07-1.02(m，3H)；ESI-MS m/z 827.00(M+Na) ⁺。

Embodiment 9 Compound II per a-9

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] urethanum

According to the method in embodiment 5, Compound II per a-2 and Vinyl chloroformate condensation purifying can be prepared Compound II per a-9.

¹H NMR(400MHz，CDCl ₃)δ9.94(brs，1H)，7.43-7.26(m，3H)，7.16(s，1H)，6.70(d，J＝15.2Hz，1H)，5.98(d，J＝16.4Hz，1H)，5.70-5.60(m，2H)，5.34(s，1H)，5.23-5.14(m，2H)，4.83-4.72(m，4H)，4.52-3.98(m，9H)，3.81-3.79(m，1H)，3.54-3.47(m，1H)，2.89-2.82(m，1H)，2.57-2.53(m，1H)，2.38-2.28(m，1H)，2.22-2.15(m，1H)，2.08-1.91(m，3H)，1.42-1.33(m，2H)，1.27-1.25(m，3H)，1.05(s，9H)；ESI-MS m/z821.00(M+Na) ⁺。

Embodiment 10 Compound II per a-10

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] benzyl carbamate

According to the method in embodiment 5, Compound II per a-2 and chloroformic acid benzyl ester condensation purifying can be prepared Compound II per a-10.

¹H NMR(400MHz，CDCl ₃)δ7.51(s，1H)，7.48-7.26(m，7H)，7.15(s，1H)，6.70(d，J＝17.2Hz，1H)，5.98(d，J＝16.0Hz，1H)，5.85-5.60(m，2H)，5.40-5.09(m，5H)，4.85-4.73(m，4H)，4.48-4.40(m，3H)，4.28-4.20(m，2H)，4.09-3.97(m，2H)，3.81-3.76(m，1H)，3.54-3.47(m，1H)，2.88-2.82(m，1H)，2.58-2.53(m，1H)，2.34-2.26(m，1H)，2.06-1.91(m，2H)，1.39-1.33(m，2H)，1.31-1.25(m，2H)，0.99(s，9H)；ESI-MS m/z882.65(M+Na) ⁺。

Embodiment 11 Compound II per a-11

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] carboxylamine ring pentyl ester

According to the method in embodiment 5, Compound II per a-2 and chloroformate cyclopentyl ester condensation purifying can be prepared Compound II per a-11.

¹H NMR(400MHz，CDCl ₃)δ7.50(s，2H)，7.30-7.26(m，2H)，7.16(s，1H)，6.70(d，J＝16.8Hz，1H)，5.98(d，J＝16.8Hz，1H)，5.80-5.67(m，2H)，5.33-5.12(m，4H)，4.83-4.72(m，4H)，4.52-4.40(m，3H)，4.31-4.25(m，2H)，4.09-3.99(m，2H)，3.81-3.77(m，1H)，3.54-3.48(m，1H)，2.89-2.82(m，1H)，2.57-2.53(m，1H)，2.36-2.26(m，1H)，2.08-1.80(m，6H)，1.59-1.50(m，4H)，1.39-1.33(m，2H)，1.10-1.05(m，2H)，1.05(s，9H)；ESI-MS m/z 861.00(M+Na) ⁺。

Embodiment 12 Compound II per a-12

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-17-[(tertiary butyl carboxamide) is amino]-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides

According to the method in embodiment 5, Compound II per a-2 and t-butylisocyanate condensation purifying can be prepared Compound II per a-12.

¹H NMR(400MHz，CDCl ₃)δ8.39(brs，1H)，8.02(d，J＝8.0Hz，1H)，7.22(d，J＝7.2Hz，1H)，7.13-7.11(m，2H)，7.19-7.07(m，2H)，6.52(d，J＝15.6Hz，1H)，5.88(d，J＝16.0Hz，1H)，5.61-5.55(m，2H)，5.37(s，1H)，5.25-5.12(m，2H)，4.85-4.20(m，10H)，3.91-3.83(m，3H)，3.40-3.38(m，1H)，2.89-2.72(m，1H)，2.59-2.50(m，1H)，2.34-2.20(m，1H)，2.10-2.04(m，2H)，1.88-1.80(m，2H)，1.33(s，9H)，1.06(s，9H)；ESI-MS m/z 848.00(M+Na) ⁺。

Embodiment 13 Compound II per a-13

N-[(1R, 17S, 20S, the 23S)-20-tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate

Be hydrolyzed after intermediate A 8 hydrogenation in embodiment 1, then can prepare Compound II per a-13 with the right fragment C7 condensation purifying.

Step one: preparation intermediate A 9-1

10%Pd/C (30mg, 20%) is added in the 10mL ethyl acetate solution of intermediate A 8 (75mg, 0.12mmol).Atmospheric hydrogenation stirs and spends the night, and obtain hydrogenated products crude product (57mg, 76.0%) after filtering and concentrating, pale yellow oil, without purifying, is directly used in saponification.

Subsequently above-mentioned hydrogenated products (57mg, 0.09mmol) is added to the THF/H of LiOH (12mg, 0.27mmol) ₂in O (5mL/2.5mL) solution, stir 1 hour.PH 1N HCl is adjusted to 4, and water layer is separated, and adds extraction into ethyl acetate twice.Saturated common salt is washed, Na ₂sO ₄drying, obtains pale yellow oil A9-1 (52mg, 93.4%) after filtering and concentrating.Without purifying, be directly used in condensation.

Step 2: prepare Compound II per a-13

According to flow process IV, by left side acid fragment A9-1 (52mg, 0.08mmol), amine fragment C7 (29mg, 0.08mmol), HATU (50mg, 0.12mmol) be dissolved in (10mL) in methylene dichloride with DiPEA (20mg, 0.16mmol) mixing.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains white powder IIa-13 (16mg, 22.9%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ10.05(brs，1H)，7.58(s，1H)，7.20(d，J＝7.6Hz，1H)，7.10-7.05(m，2H)，5.80-5.72(m，1H)，5.45-5.38(m，2H)，5.27-5.13(m，2H)，4.79-4.65(m，4H)，4.53-4.29(m，4H)，3.81-3.72(m，2H)，3.55-3.46(m，3H)，2.90-2.82(m，1H)，2.75-2.68(m，1H)，2.55-2.50(m，2H)，2.35-2.20(m，1H)，2.09-1.80(m，4H)，1.59-1.50(m，2H)，1.45(s，9H)，1.04(s，9H)，1.00-0.96(m，2H)；ESI-MS m/z 851.00(M+Na) ⁺。

Embodiment 14 Compound II per a-14

N-[(1R, 17S, 20S, the 23S)-20-tertiary butyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate

Remove Boc by after the intermediate C6 hydrogenation in embodiment 1, then can prepare Compound II per a-13 with left side Segment A 9-1 condensation purifying.

Step one: prepare intermediate C7-1

10%Pd/C (80mg, 20%) is added in the 10mL ethyl acetate solution of intermediate C6 (200mg, 0.65mmol).Atmospheric hydrogenation stirs and spends the night, and filtering and concentrating, obtains hydrogenated products (120mg, 58.0%), white solid after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ9.68(brs，1H)，5.08(brs，1H)，2.98-2.92(m，1H)，1.60-1.56(m，1H)，1.47(s，9H)，1.45-1.39(m，4H)，1.35-1.29(m，2H)，1.11-1.06(m，2H)，1.01(t，J＝7.2Hz，3H).

According to flow process III, above-mentioned hydrogenated products intermediate (40mg, 0.12mmol) is dissolved in methylene dichloride (5mL), drips trifluoroacetic acid (1mL, 20%) and stir 2 hours afterwards.Brown oil C7-1 (42mg, 97.3%) is obtained after concentrated.Without purifying, be directly used in condensation.

Step 2: prepare Compound II per a-14

According to flow process IV, by left side acid fragment A9-1 (80mg, 0.13mmol), amine fragment C7-1 (42mg, 0.13mmol), HATU (80mg, 0.20mmol) and DiPEA (34mg, 0.26mmol) mixing is dissolved in (10mL) in methylene dichloride.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains white powder IIa-14 (4.3mg, 15.7%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ7.52(brs，1H)，7.33(brs，1H)，7.21(t，J＝7.6Hz，1H)，7.11(d，J＝7.6Hz，1H)，7.05(d，J＝7.6Hz，1H)，5.43-5.40(m，1H)，5.37(s，1H)，4.79-4.67(m，4H)，4.54-4.44(m，2H)，4.35-4.27(m，2H)，3.78-3.75(m，2H)，3.54-3.50(m，2H)，3.46-3.40(m，1H)，2.93-2.90(m，1H)，2.69-2.62(m，1H)，2.60-2.54(m，1H)，2.40-2.33(m，1H)，2.22-2.18(m，1H)，2.00-1.90(m，2H)，1.64-1.60(m，2H)，1.54-1.50(m，2H)，1.44(s，9H)，1.36-1.30(m，2H)，1.14-1.10(m，2H)，1.04(s，9H)，0.96-0.93(m，3H)；ESI-MS m/z 853.00(M+Na) ⁺。

Embodiment 15 Compound II per a-15

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

According to flow process IV, by left side acid fragment A9 (98mg, 0.16mmol), amine fragment C7-1 (56mg, 0.16mmol), HATU (92mg, 0.24mmol) and DiPEA (342mg, 0.32mmol) mixing is dissolved in (10mL) in methylene dichloride.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains white powder IIa-15 (56mg, 42.4%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ7.48(brs，1H)，7.30-7.24(m，2H)，7.16(d，J＝6.4Hz，1H)，6.77(d，J＝16.4Hz，1H)，6.05(d，J＝16.4Hz，1H)，5.58(d，J＝7.2Hz，1H)，5.29(s，1H)，4.89-4.85(m，1H)，4.77-4.71(m，3H)，4.46-4.29(m，6H)，4.09-4.00(m，2H)，3.76-3.73(m，1H)，3.46-3.40(m，1H)，2.93-2.90(m，1H)，2.57-2.50(m，1H)，2.40-2.33(m，1H)，1.63-1.52(m，4H)，1.49(s，9H)，1.36-1.18(m，5H)，1.05(s，9H)，0.96(t，J＝8.0Hz，3H)；ESI-MS m/z 851.00(M+Na) ⁺。

Embodiment 16 Compound II per a-16

N-[(1R, 12E, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

According to the IIa-1 method preparing compound, change Segment A 4---N-Boc-L-Terleu into N-Boc-L-2-Cyclohexylglycine and can prepare Compound II per a-16.

¹H NMR(400MHz，CDCl ₃)δ10.09(brs，1H)，7.30-7.27(m，2H)，7.20-7.16(m，2H)，6.76(d，J＝16.0Hz，1H)，6.04(d，J＝16.0Hz，1H)，5.75-5.70(m，1H)，5.55-5.50(m，1H)，5.33(s，1H)，5.25-5.11(m，2H)，4.84-4.74(m，4H)，4.42-4.29(m，5H)，4.15-4.10(m，1H)，4.04-4.00(m，1H)，3.95-3.85(m，1H)，3.79-3.75(m，1H)，3.52-3.48(m，1H)，2.92-2.85(m，1H)，2.53-2.48(m，1H)，2.45-2.40(m，1H)，2.02-1.95(m，2H)，1.76-1.70(m，4H)，1.46(s，9H)，1.36-0.98(m，12H)；ESI-MS m/z 875.00(M+Na) ⁺。

Embodiment 17 Compound II per a-17

N-[(1R, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate

Compound II per a-17 is prepared according to the method preparing Compound II per a-13.

¹H NMR(400MHz，CDCl ₃)δ10.02(brs，1H)，7.65(brs，1H)，7.40(brs，1H)，7.23(t，J＝7.6Hz，1H)，7.11(d，J＝7.6Hz，1H)，7.05(d，J＝7.6Hz，1H)，5.72-5.63(m，1H)，5.50(d，J＝7.6Hz，1H)，5.41(s，1H)，5.25-5.12(m，2H)，4.79-4.65(m，3H)，4.56-4.48(m，2H)，4.39-4.32(m，3H)，3.83-3.80(m，1H)，3.73-3.71(m，1H)，3.56-3.53(m，1H)，3.46-3.43(m，2H)，2.90-2.86(m，1H)，2.74-2.67(m，1H)，2.58-2.50(m，2H)，2.42-2.36(m，1H)，2.07-2.00(m，1H)，1.95-1.92(m，1H)，1.82-1.65(m，6H)，1.44(s，9H)，1.37-1.31(m，4H)，1.21-1.15(m，3H)，1.08-0.98(m，5H)；ESI-MS m/z 877.00(M+Na) ⁺。

Embodiment 18 Compound II per a-18

N-[(1R, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate

Compound II per a-18 is prepared according to the method preparing Compound II per a-14.

¹H NMR(400MHz，CDCl ₃)δ10.06(brs，1H)，7.55(brs，1H)，7.43(brs，1H)，7.23(t，J＝7.6Hz，1H)，7.11(d，J＝7.6Hz，1H)，7.05(d，J＝7.6Hz，1H)，5.49(d，J＝7.2Hz，1H)，5.40(s，1H)，4.78-4.65(m，3H)，4.56-4.47(m，2H)，4.39-4.31(m，3H)，3.83-3.80(m，1H)，3.73-3.71(m，1H)，3.56-3.54(m，1H)，3.46-3.43(m，2H)，2.96-2.90(m，1H)，2.75-2.68(m，1H)，2.58-2.48(m，2H)，2.42-2.36(m，1H)，2.31-2.17(m，2H)，1.79-1.58(m，8H)，1.44(s，9H)，1.37-1.30(m，4H)，1.22-1.15(m，3H)，1.13-1.03(m，5H)，0.98(t，J＝7.2Hz，3H)；ESI-MS m/z 879.00(M+Na) ⁺。

Embodiment 19 Compound II per a-19

N-[(1R, 12E, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

Compound II per a-19 is prepared according to the method preparing Compound II per a-15.

¹H NMR(400MHz，CDCl ₃)δ10.02(brs，1H)，7.32-7.25(m，1H)，7.17-7.16(m，1H)，6.77(d，J＝16.4Hz，1H)，6.05(d，J＝16.4Hz，1H)，5.53(d，J＝6.4Hz，1H)，5.35(s，1H)，4.89-4.86(m，1H)，4.78-4.73(m，3H)，4.45-4.30(m，3H)，4.15-4.11(m，1H)，4.05-4.03(m，1H)，3.78-3.76(m，1H)，3.55-3.51(m，1H)，2.99-2.91(m，1H)，2.58-2.52(m，1H)，2.48-2.41(m，1H)，1.81-1.57(m，8H)，1.48(s，9H)，1.41-1.32(m，5H)，1.22-1.15(m，3H)，1.09-1.03(m，4H)，0.98(t，J＝7.2Hz，3H)；ESI-MS m/z 879.00(M+Na) ⁺。

Embodiment 20 Compound II per a-20

N-[(1R, 12E, 18S, 21S, 24S)-21-the tertiary butyl-24-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,19,22-trioxy--2,15-dioxa-4,20,23-tri-azepine Fourth Ring [21.2.1.1 ^4,7.0 ^6,11] 27 yuan-6,8,10,12-tetraene-18-position] t-butyl carbamate

Change raw material N-Boc-L-serine methylester into N-Boc-L-homoserine methylether according to the method preparing Compound II per a-1 and can prepare Compound II per a-20.

¹H NMR(400MHz，CDCl ₃)δ7.57(brs，1H)，7.28-7.25(m，2H)，7.16(d，J＝7.2Hz，1H)，6.52(d，J＝18.0Hz，1H)，6.04(d，J＝18.0Hz，1H)，5.67-5.55(m，1H)，5.46-5.40(m，2H)，5.34-5.12(m，2H)，4.75-4.61(m，6H)，4.45-4.40(m，2H)，4.25-4.22(m，2H)，4.06-4.00(m，2H)，3.93-3.90(m，1H)，3.50-3.47(m，2H)，2.89-2.85(m，1H)，2.57-2.50(m，1H)，2.38-2.31(m，1H)，2.17-2.08(m，2H)，1.85-1.80(m，1H)，1.43(s，9H)，1.40-1.20(m，6H)，1.06(s，9H)；ESI-MS m/z 863.05(M+Na) ⁺。

Embodiment 21 Compound II per a-21

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2-oxa--15-thia-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

Change raw material N-Boc-L-serine methylester into N-Boc-L-acthiol-J according to the method preparing Compound II per a-1 and can prepare Compound II per a-21.

¹H NMR(400MHz，CDCl ₃)δ9.96(brs，1H)，7.64(brs，1H)，7.39-7.25(m，2.7H)，7.18-7.16(m，1.3H)，6.19(d，J＝15.6Hz，1H)，5.75-5.65(m，1H)，5.54(brs，1H)，5.32-5.13(m，2H)，5.07(d，J＝7.2Hz，1H)，4.79-4.68(m，4H)，4.56-4.53(m，3H)，4.39(d，J＝11.2Hz，1H)，4.26(brs，1H)，3.80-3.76(m，1H)，3.13(d，J＝7.6Hz，2H)，3.03-3.00(m，1H)，2.93-2.87(m，1H)，2.50-2.43(m，1H)，2.37-2.30(m，1H)，2.01-1.94(m，1H)，1.48(s，9H)，1.45-1.39(m，2H)，1.39-1.27(m，4H)，1.05(s，9H)；ESI-MS m/z 865.00(M+Na) ⁺。

Embodiment 22 Compound II per a-22

N-[(1R, 12E, 17S, 20S, 23S)-20-cyclopentyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate

According to the IIa-1 method preparing compound, change Segment A 4---N-Boc-L-Terleu into N-Boc-L-2-cyclopentylglycine and can prepare Compound II per a-22.

¹H NMR(400MHz，CDCl ₃)δ10.00(brs，1H)，7.66-7.57(m，1H)，7.33-7.23(m，2H)，7.17-7.13(m，2H)，6.69-6.53(m，1H)，6.01-5.78(m，3H)，5.52-5.10(m，4H)，4.85-4.67(m，4H)，4.51-4.18(m，6H)，3.90-3.81(m，2H)，3.61-3.34(m，1H)，2.92-2.85(m，1H)，2.56-2.50(m，1H)，2.40-2.36(m，1H)，2.09-1.99(m，2H)，1.78-1.72(m，2H)，1.60-1.55(m，4H)，1.49(s，9H)，1.46-1.35(m，4H)，1.13-1.01(m，4H)；ESI-MS m/z 861.00(M+Na) ⁺。

Embodiment 23 Compound II per b-1

N-[(3R, 5S, 8S, 11S, 15E)-8-the tertiary butyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate

Synthesize according to the method for flow process II, III, IV.

Intermediate B 2:(2S, 4R)-4-(7-bromo-6-methoxyl group isoquinoline give repeated exhortations-1-position oxygen base) tetramethyleneimine-1-t-butyl formate-2-methyl-formiate

According to flow process I, raw material A 2 (466mg, 2.01mmol) is dissolved in DMSO (10mL), then adds in batches ^tbuOK (616mg, 5.49mmol).Stirring at room temperature, after 20 minutes, slowly drips the DMSO solution of B1 (prepared by reference literature, WO2008/051475) (500mg, 1.83mmol), drips Bi Jixu and stirs 2 hours.Add 2N HCl cancellation react and regulate pH=1-2, be then extracted with ethyl acetate three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, obtains acid crude A3 (600mg, 70.6%) after filtering and concentrating.Without purifying, continue to drop into next step reaction.

Above-mentioned acid crude (4.0g, 8.62mmol) is dissolved in DMF (50mL), then adds K ₂cO ₃(2.38g, 17.24mmol).Slow dropping CH ₃i (1.59g, 11.2mmol), drips Bi Jixu and stirs 1 hour.Then reaction solution is poured into water, is extracted with ethyl acetate three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains intermediate B 2 (3.7g, 89.4%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ8.22(s，1H)，7.81-7.79(m，1H)，7.00-6.98(m，1H)，6.89(s，1H)，5.67-5.64(m，1H)，4.51-4.44(m，1H)，3.89-3.87(m，5H)，3.69(s，3H)，2.53-2.45(m，1H)，2.29-2.22(m，1H)，1.37(s，9H).

Intermediate B 3:(2S, 4R)-4-(7-vinyl-6-methoxyl group isoquinoline give repeated exhortations-1-position oxygen base) tetramethyleneimine-1-t-butyl formate-2-methyl-formiate

Above-mentioned bromide B2 (1.1g, 2.29mmol) is dissolved in toluene (25mL), then adds Bu ₃snCH=CH ₂(1.09g, 3.44mmol) and Pd (PPh ₃) ₄(264mg, 0.229mmol).Reflux 5 hours under nitrogen protection.Concentrated, obtain intermediate B 3 (0.84g, 86.0%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ8.18(s，1H)，7.87-7.84(m，1H)，7.11-7.09(m，2H)，6.98(s，1H)，5.93(d，J＝17.2Hz，1H)，5.77-5.75(m，1H)，5.4(d，J＝11.6Hz，1H)，4.62-4.51(m，1H)，3.97-3.93(m，5H)，3.78(s，3H)，2.66-2.58(m，1H)，2.39-2.35(m，1H)，1.44(s，9H).

Intermediate B 4:(2S; 4R)-1-((S)-2-(t-butoxycarbonyl amino)-3,3-dimethylbutanoyl)-4-(6-methoxyl group-7-vinyl isoquinoline give repeated exhortations-1-position oxygen base) tetramethyleneimine-2-methyl-formiate

According to flow process II, raw material B3 (840mg, 1.96mmol) is dissolved in methylene dichloride (10mL), drip trifluoroacetic acid (3mL) under ice bath, stirring at room temperature is after 3 hours, and reaction terminates, concentrated, slowly drip aqueous sodium carbonate, extraction into ethyl acetate three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, obtains brown oil (550mg, 86%) after filtering and concentrating.Without purifying, directly drop into next step reaction.

By N-Boc-L-Terleu (636mg, 2.75mmol), de-Boc crude product (900mg, 2.75mmol), HATU (1.36g, 3.57mmol) and DiPEA (460mg, 3.57mmol) mixing is dissolved in methylene dichloride (15mL).Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains brown oil B4 (810mg, 54.5%) after column chromatography.

ESI-MS m/z 564.0(M+H) ⁺。

Intermediate B 5:(2S; 4R)-1-((S)-2-((S)-3-(allyloxy)-2-(t-butoxycarbonyl amino) propionic acid amide)-3,3-dimethylbutanoyl)-4-(6-methoxyl group-7-vinyl isoquinoline give repeated exhortations-1-position oxygen base) tetramethyleneimine-2-methyl-formiate

According to flow process I, by raw material B4 (810mg, 1.5mmol) be dissolved in methylene dichloride (12mL), then drip in trifluoroacetic acid (2.5mL), stirring at room temperature is after 2 hours, and reaction terminates, and adds water and methylene dichloride after concentrated, and regulate pH to 12 with 2N NaOH, extract three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating must take off Boc crude product.Without purifying, directly drop into next step reaction.

By amino acid A6 (363mg, 1.48mmol), de-Boc crude product (652mg, 1.48mmol), HATU (732mg, 1.92mmol) and DiPEA (248mg, 1.92mmol) mixing is dissolved in (20mL) in methylene dichloride.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains pale yellow oil B5 (830mg, 84.07%) after column chromatography.

ESI-MS m/z 691.50(M+Na) ⁺。

Intermediate B 6:N-[(3R, 5S, 8S, 11S, 15E)-8-tertiary butyl-5-(methanoyl methyl)-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate

According to flow process I; under nitrogen protection; raw material B5 (830mg, 1.244mmol) is dissolved in methylene dichloride (340mL), then adds Zhan catalyst (90mg; 0.124mmol); after stirred overnight at room temperature, reaction terminates, and adds 0.1mL DMSO cancellation reaction; after concentrated, column chromatography obtains brown oil B6 (540mg, 68.0%).

ESI-MS m/z 663.50(M+Na) ⁺。

Intermediate B 7:N-[(3R, 5S, 8S, 11S, 15E)-8-tertiary butyl-5-(methanoyl hydroxyl)-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate

According to flow process I, intermediate B 7 (170mg, 0.266mmol) is added in MeOH/THF/H2O (4mL/4mL/1mL) solution of LiOH (67mg, 1.6mmol), stirs 2 hours.After concentrated, pH 1N HCl is adjusted to 4, water layer is separated, and adds extraction into ethyl acetate twice.Saturated common salt is washed, Na ₂sO ₄drying, obtains pale yellow oil A6 (160mg, 100%) after filtering and concentrating.Without purifying, be directly used in condensation.

Compound II per b-1

According to flow process IV, by left side acid fragment B7 (160mg, 0.255mmol), amine fragment C7 (64mg, 0.280mmol), HATU (126mg, 0.331mmol) and DiPEA (43mg, 0.331mmol) mixing is dissolved in (6mL) in methylene dichloride.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains white powder IIb-1 (26mg, 12.0%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ8.47(s，1H)，7.87(d，J＝5.6Hz，1H)，7.52(d，J＝6.0Hz，1H)，7.36(s，1H)，7.12(d，J＝5.6Hz，1H)，6.94(s，1H)，6.90(d，J＝15.6Hz，1H)，6.35-6.28(m，1H)，5.71-5.61(m，3H)，5.22(d，J＝16.8Hz，1H)，5.11(d，J＝10.8Hz，1H)，4.60-4.53(m，2H)，4.45-4.40(m，1H)，4.34-4.26(m，2H)，4.04-3.89(m，6H)，2.90-2.86(m，1H)，2.78-2.73(m，1H)，2.47-2.41(m，1H)，2.05-1.99(m，2H)，1.93-1.90(m，1H)，1.45(s，9H)，1.42-1.38(m，2H)，1.08(s，9H)1.02-0.98(m，2H)；ESI-MS m/z 861.00(M+Na) ⁺。

Embodiment 24 Compound II per b-2

N-[(3R, 5S, 8S, 11S, 15E)-8-the tertiary butyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 17 (25), 18,20 (24), 21-pentaene-11-positions] t-butyl carbamate

Be hydrolyzed after intermediate B 6 hydrogenation in embodiment 23, then can prepare Compound II per b-2 with the right fragment C7 condensation purifying.

Step one: preparation intermediate B 7-1

10%Pd/C (80mg, 10%Pd/C) is added in the 30mL ethyl acetate solution of intermediate B 6 (170mg, 0.271mmol).Atmospheric hydrogenation stirs 3 hours, obtains hydrogenated products crude product, pale yellow oil after filtering and concentrating, without purifying, is directly used in saponification.

Subsequently above-mentioned hydrogenated products crude product is added to the MeOH/THF/H of LiOH (64mg, 1.527mmol) ₂in O (4mL/4mL/1mL) solution, stir 2 hours.PH 1N HCl is adjusted to 4, and water layer is separated, and adds extraction into ethyl acetate twice.Saturated common salt is washed, Na ₂sO ₄drying, obtains pale yellow oil B7-1 (150mg, 96.0%) after filtering and concentrating.Without purifying, be directly used in condensation.

Step 2: prepare Compound II per b-2

According to flow process IV, by left side acid fragment B7-1 (150mg, 0.238mmol), amine fragment C7 (118mg, 0.309mmol), HATU (126mg, 0.331mmol) and DiPEA (46mg, 0.357mmol) mixing is dissolved in (6mL) in methylene dichloride.Room temperature for overnight, adds water and methylene dichloride, extracts three times.Organic phases washed with water, saturated common salt is washed, Na ₂sO ₄drying, filtering and concentrating, obtains white powder IIb-2 (80mg, 52.5%) after column chromatography.

¹H NMR(400MHz，CDCl ₃)δ7.86(d，J＝6.0Hz，1H)，7.82(s，1H)，7.60(brs，1H)，7.30(d，J＝8.8Hz，1H)，7.13(d，J＝6.0Hz，1H)，6.94(s，1H)，5.73(brs，1H)，5.69-5.64(m，1H)，5.48(d，J＝7.6Hz，1H)，5.23(d，J＝16.8Hz，1H)，5.12(d，J＝10.4Hz，1H)，4.85(d，J＝8.8Hz，1H)，4.50-4.47(m，2H)，4.40-4.37(m，2H)，3.92-3.87(m，4H)，3.64-3.60(m，2H)，3.37-3.30(m，2H)，2.87-2.72(m，3H)，2.45-2.39(m，1H)，2.06-1.99(m，1H)，1.95-1.89(m，3H)，1.44(s，9H)，1.40-1.36(m，2H)，1.06(s，9H)，1.02-0.98(m，4H)；ESI-MS m/z 863.00(M+Na) ⁺。

Embodiment 25 Compound II per b-3

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate

According to the method for synthetic compound IIb-1, change Segment A 4---N-Boc-L-Terleu into N-Boc-L-2-Cyclohexylglycine and can prepare Compound II per b-3.

¹H NMR(400MHz，CDCl ₃)δ8.40(s，1H)，7.88(d，J＝6.0Hz，1H)，7.26(brs，1H)，7.16(brs，1H)，7.13(d，J＝6.0Hz，1H)，6.94(s，1H)，6.90(d，J＝16.0Hz，1H)，6.31-6.24(m，1H)，5.75-5.67(m，2H)，5.55(d，J＝4.2Hz，1H)，5.22(d，J＝16.4Hz，1H)，5.11(d，J＝11.2Hz，1H)，4.55-4.48(m，2H)，4.41-4.36(m，1H)，4.31-4.24(m，2H)，4.11-4.06(m，1H)，3.94(s，3H)，3.92-3.87(m，2H)，3.57-3.53(m，1H)，2.92-2.85(m，1H)，2.73-2.68(m，1H)，2.54-2.47(m，1H)，2.01-1.95(m，2H)，1.86-1.67(m，1H)，1.45(s，9H)，1.37-1.29(m，4H)，1.15-0.99(m，4H)；ESI-MS m/z887.00(M+Na) ⁺。

Embodiment 26 Compound II per b-4

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 17 (25), 18,20 (24), 21-pentaene-11-positions] t-butyl carbamate

According to the method for synthetic compound IIb-2, change Segment A 4---N-Boc-L-Terleu into N-Boc-L-2-Cyclohexylglycine and can prepare Compound II per b-4.

¹H NMR(400MHz，CDCl ₃)δ7.87(d，J＝6.0Hz，1H)，7.82(s，1H)，7.63(brs，1H)，7.39(brs，1H)，7.13(d，J＝6.0Hz，1H)，6.94(s，1H)，5.80(brs，1H)，5.72-5.64(m，1H)，5.51(d，J＝7.6Hz，1H)，5.23(d，J＝17.2Hz，1H)，5.12(d，J＝10.4Hz，1H)，4.72-4.68(m，1H)，4.54(d，J＝11.6Hz，1H)，4.42-4.38(m，2H)，3.94-3.92(m，1H)，3.92(s，3H)，3.63(d，J＝4.0Hz，2H)，3.38-3.29(m，2H)，2.89-2.83(m，2H)，2.77-2.71(m，2H)，2.50-2.43(m，1H)，2.03-1.97(m，2H)，1.93-1.85(m，4H)，1.75-1.65(m，4H)，1.43(s，9H)，1.32-1.27(m，4H)，1.07-0.99(m，4H)；ESI-MS m/z 889.00(M+Na) ⁺。

Embodiment 27 Compound II per b-5

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate

According to the method for synthetic compound IIb-1, fragment C7 is changed into fragment C7-1 and can prepare Compound II per b-5.

¹H NMR(400MHz，CDCl ₃)δ10.02(s，1H)，8.43(s，1H)，7.91(d，J＝6.0Hz，1H)，7.19(d，J＝6.0Hz，1H)，7.07(brs，1H)，6.98(s，1H)，6.92(d，J＝16.0Hz，1H)，6.32-6.24(m，1H)，5.78(brs，1H)，5.60(d，J＝8.0Hz，1H)，4.60-4.48(m，2H)，4.47-4.30(m，3H)，4.11-4.06(m，1H)，3.97(s，3H)，3.95-3.91(m，2H)，3.56-3.50(m，1H)，2.94-2.90(m，1H)，2.80-2.73(m，1H)，2.60-2.50(m，1H)，1.93-1.65(m，10H)，1.48(s，9H)，1.32-1.27(m，6H)，1.16-1.06(m，4H)，0.95(t，J＝7.2Hz，3H)；ESI-MS m/z 888.75(M+Na) ⁺。

Embodiment 28 Compound II per b-6

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 17 (25), 18,20 (24), 21-pentaene-11-positions] t-butyl carbamate

According to the method for synthetic compound IIb-4, fragment C7 is changed into fragment C7-1 and can prepare Compound II per b-6.

¹H NMR(400MHz，CDCl ₃)δ10.06(s，1H)，7.90(d，J＝6.0Hz，1H)，7.86(s，1H)，7.18(d，J＝6.0Hz，1H)，6.98(s，1H)，5.86(brs，1H)，5.51(brs，1H)，4.70-4.60(m，1H)，4.58(d，J＝10.8Hz，1H)，4.50-4.42(m，2H)，3.99-3.97(m，1H)，3.94(s，3H)，3.63(d，J＝4.0Hz，2H)，3.37-3.30(m，2H)，2.94-2.89(m，2H)，2.78-2.71(m，2H)，2.54-2.48(m，1H)，2.00-1.50(m，10H)，1.45(s，9H)，1.39-1.20(m，8H)，1.07-0.99(m，4H)，0.96(t，J＝7.2Hz，3H)；ESI-MS m/z 866.69(M-H) ^-。

Embodiment 29 Compound II per b-7

(1R, 2S)-1-[(3R, 5S, 8S, 11S, 15E)-11-{ [(tertbutyloxycarbonyl] amino-8-cyclohexyl-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23 3 azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-5-carbamyls]-2-vinyl cyclopropyl-1-carboxylic acid

According to the method for synthetic compound IIb-1, after fragment C7 being changed into fragment C5 condensation, hydrolysis can prepare Compound II per b-7.

¹H NMR(400MHz，CDCl ₃)δ8.27(s，1H)，7.83(d，J＝5.2Hz，1H)，7.50(brs，1H)，7.32(d，J＝6.0Hz，1H)，7.08(d，J＝5.2Hz，1H)，6.89(s，1H)，6.82(d，J＝16.0Hz，1H)，6.21-6.15(m，1H)，5.71-5.64(m，2H)，5.52(d，J＝7.2Hz，1H)，5.22-5.04(m，2H)，4.60-4.48(m，2H)，4.42(d，J＝9.6Hz，1H)，4.23-4.20(m，1H)，4.11(d，J＝6.0Hz，2H)，3.88(s，3H)，3.80-3.70(m，2H)，3.43(t，J＝7.2Hz，1H)，2.80-2.73(m，1H)，2.60-2.50(m，1H)，1.98-1.88(m，2H)，1.73-1.50(m，8H)，1.45(s，9H)，1.18-1.01(m，4H)；ESI-MS m/z 760.00(M-H) ^-。

The biological activity test of embodiment 30 compound

The ability that the compound of formula I suppresses HCV to copy is proved by following experiment in vitro.

HCV replicon (Replicon) is tested

DMEM substratum: obtain by adding the preparation of 10%FBS, 2mM L-glutamic acid (Life Technologies #25030-024), penicillin (100IU/ml)/Streptomycin sulphate (100g/ml) (Life Technologies #15140-114) and 1x non-essential amino acid (Life Technologies #11140-035) to DMEM (Life Technologies #41965-039).

Perfect medium is prepared from after being mixed by foetal calf serum (FBS), DMEM substratum and G418.Above-mentioned perfect medium is poured in Tissue Culture Dish and is placed in 37 DEG C of CO ₂preheating in constant incubator.Afterwards the huh-7 cell (V.Lohmann etc., Science, 285, (1999) 110-113) at hepatitis C virus (HCV) replicon cultivated is taken out from 37 DEG C of incubators.Carefully suck the substratum in culture dish, and clean cell with phosphoric acid buffer (PBS).1ml 0.25% trypsin Tyrpsin is added after removing above-mentioned scavenging solution)/0.02%EDTA solution.Clean cell with above-mentioned trypsinase/EDTA solution and guarantee that all cells is all cleaned.In 37 DEG C of incubators, 3-5 minute is hatched after sucking trypsinase/EDTA solution.By inverted microscope observation of cell form, to culturing cell depart from wall of container and be separated from each other high-visible.

The perfect medium adding 3ml preincubate is to culture dish and by above-mentioned cell suspension.Cell quantity is counted with hemometer (Hematometer).Cell density is adjusted to 100,000/ml by the perfect medium adding proper volume.The cell suspending liquid adding 100 μ l in each hole in 96 orifice plates makes every porocyte concentration be 10,000. 96 orifice plates are placed in 37 DEG C of 5%CO ₂24 hours are hatched in constant incubator.

The preparation of test compounds solution.The dilution of all compounds all should aseptically be carried out.

Compound storage liquid can be prepared before on-test, by testing compound be dissolved in 100%DMSO to final concentration be 2mM.

96 orifice plates cultivate and terminate before by above-claimed cpd storage liquid cell culture medium to 100 times (100 × solution) of required final detectable level.

By 96 orifice plates from 37 DEG C of 5%CO ₂shift out in constant incubator and use inverted microscope observation of cell form.

In cell cultures stink cupboard, add in 96 orifice bores by 1 μ l 100 × compound solution, in system, DMSO final concentration is 1%.96 orifice plates adding compound solution are placed in CO ₂cultivate 48 hours in incubator.

30 μ L Stead-Glo Luciferase Assay System (purchased from Pu Luomaige Promega) reagent are added in each hole of 96 orifice plates, by the mixing in 5 minutes of jog on shaking table to guarantee the complete cracking of cell.Read fluorescence values with Envision (purchased from perkin elmer instrument Perkin Elmer), arranging integral time was 2 seconds.

Record also obtains following data after analytical data.

Test-results (the EC on replicon 1b of compound in embodiment ₅₀value) list in table 1, wherein A represents its EC ₅₀value is less than 10nM, and B represents its EC ₅₀value is between 10nM to 50nM, and C represents its EC ₅₀value is between 50nM to 500nM, and D represents its EC ₅₀value is between 500nM to 20uM, and E represents its EC ₅₀value is greater than 20uM.

Compound	EC50	Compound	EC50	Compound	EC50
						IIa-1	A	IIa-2	B	IIa-3	C
IIa-4	D	IIa-5	C	IIa-6	C
						IIa-7	C	IIa-8	B	IIa-9	B
IIa-10	A	IIa-11	A	IIa-12	A
						IIa-13	B	IIa-14	C	IIa-15	A
IIa-16	A	IIa-17	A	IIa-18	B
						IIa-19	A	IIa-20	A	IIa-21	B
IIa-22	A	IIb-1	A	IIb-2	A
						IIb-3	A	IIb-4	A	IIb-5	B
IIb-6	C	IIb-7	D	MK7009	A

Result shows, the invention provides the compound with strong HCV-Ab IgG virus activity of a class formation novelty.

MK7009 is the compound in documents WO 2007/015787, and structure is as follows:

The medicine of embodiment 31 compound is for property test

The medicine of the compound of formula I is proved by the test of hepatomicrosome metabolic stability for character:

1, buffer: buffer A: 1.0L 0.1M potassium phosphate buffer (containing 1.0mM EDTA); Buffer B: 1.0L 0.1M dipotassium hydrogen phosphate damping fluid (containing 1.0mMEDTA); Damping fluid C:0.1M potassium phosphate buffer (containing 1.0mM EDTA), pH 7.4, joins in 700mL buffer B by buffer A, stop when pH reaches 7.4.

2, compound administration solution: 500 μMs of solution: 10 μ L 10mM DMSO storage liquid are joined in 190 μ L ACN; Give drug solns (being dissolved in hepatomicrosome, hepatomicrosome final concentration 0.75mg/mL) for 1.5 μMs: 1.5 μ L, 500 μMs of solution and 18.75 μ L 20mg/mL hepatomicrosomes are joined in 479.75uL damping fluid C.

3, NADPH solution (6mM is dissolved in damping fluid C).

4,30L 1.5 μMs is joined the position being set to different time points in 96 orifice plates to drug solns.37 DEG C of preheatings 10 minutes.

5,15L NADPH solution (6mM) is joined the position being set to 45 minutes points, and start timing.

6, at 30 minutes, 15 minutes, 5 minutes, 15LNADPH solution (6mM) is joined the position of corresponding time point.

7, at the end of cultivating (0 minute), 135 μ L ACN (containing interior mark) are joined in the position being set to all time points.Then 15L NADPH solution (6mM) is joined the position being set to 0 minute.

8, centrifugal: centrifugal 10 minutes of 3220 × g.

9, take out 50 μ L supernatant liquors, mix with 50 μ L ultrapure waters (Millipore), sample presentation is analyzed to LC/MS.

In embodiment, the test-results of compound is listed in the table below:

Compound	t _1/2(minute)
		IIa-1	198
IIa-16	193
		IIb-3	230
MK7009	26

Result shows, what the invention provides a class formation novelty has the HCV-Ab IgG virus compound of outstanding medicine for character.

Above-mentioned example, only for technical conceive of the present invention and feature are described, its object is to person skilled in the art can be understood content of the present invention and implement according to this, can not limit the scope of the invention with this.All equivalent transformations of doing according to spirit of the present invention or modification, all should be encompassed within protection scope of the present invention.

Claims

1. compound or its pharmacy acceptable salt, is characterized in that, described compound is selected from:

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate;

(1R, 12E, 17S, 20S, the 23S)-17-amino-20-tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy-s-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides;

N-[(1R, 12E, 17R, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate;

(1R, 12E, 17R, 20S, the 23S)-17-amino-20-tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy-s-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides;

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy--17-C-pyrazine-2-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17,23---amide group;

(1R, 12E, 17R, 20S, 23S)-20-the tertiary butyl-23-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy--17-C-pyrazine-2-2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17,23-diamide bases;

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-17-acetylaminohydroxyphenylarsonic acid 3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides;

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-17-methanesulfonamido-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides;

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] urethanum;

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] benzyl carbamate;

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] carboxylamine ring pentyl ester;

(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-17-[(tertiary butyl carboxamide) is amino]-N-[(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl]-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-23-methane amides;

N-[(1R, 17S, 20S, the 23S)-20-tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate;

N-[(1R, 17S, 20S, the 23S)-20-tertiary butyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate;

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate;

N-[(1R, 12E, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate;

N-[(1R, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate;

N-[(1R, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10-triolefin-17-positions] t-butyl carbamate;

N-[(1R, 12E, 17S, 20S, 23S)-20-cyclohexyl-23-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate;

N-[(1R, 12E, 18S, 21S, 24S)-21-the tertiary butyl-24-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,19,22-trioxy--2,15-dioxa-4,20,23-tri-azepine Fourth Ring [21.2.1.1 ^4,7.0 ^6,11] 27 yuan-6,8,10,12-tetraene-18-position] t-butyl carbamate;

N-[(1R, 12E, 17S, 20S, 23S)-20-the tertiary butyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2-oxa--15-thia-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate;

N-[(1R, 12E, 17S, 20S, 23S)-20-cyclopentyl-23-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-3,18,21-trioxy--2,15-dioxa-4,19,22-tri-azepine Fourth Ring [20.2.1.1 ^4,7.0 ^6,11] 20 hexa-atomic-6,8,10,12-tetraene-17-positions] t-butyl carbamate;

N-[(3R, 5S, 8S, 11S, 15E)-8-the tertiary butyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate;

N-[(3R, 5S, 8S, 11S, 15E)-8-the tertiary butyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 17 (25), 18,20 (24), 21-pentaene-11-positions] t-butyl carbamate;

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate;

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2S)-1-[(cyclopropyl sulfuryl) carbamyl]-2-vinyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 17 (25), 18,20 (24), 21-pentaene-11-positions] t-butyl carbamate;

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-11-positions] t-butyl carbamate;

N-[(3R, 5S, 8S, 11S, 15E)-8-cyclohexyl-5-{ [(1R, 2R)-1-[(cyclopropyl sulfuryl) carbamyl]-2-ethyl cyclopropyl] carbamyl }-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23-tri-azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 17 (25), 18,20 (24), 21-pentaene-11-positions] t-butyl carbamate;

(1R, 2S)-1-[(3R, 5S, 8S, 11S, 15E)-11-{ [(tertbutyloxycarbonyl] amino-8-cyclohexyl-18-methoxyl group-7,10-dioxo-2,13-dioxa-6,9,23 3 azepine Fourth Ring [15.6.2.1 ^3,6.0 ^20,24] 20 hexa-atomic-1 (23), 15,17 (25), 18,20 (24), 21-six alkene-5-carbamyls]-2-vinyl cyclopropyl-1-carboxylic acid.

2. a medicinal compositions, is characterized in that, described composition contains compound according to claim 1 or its pharmacy acceptable salt of pharmaceutically acceptable carrier and pharmaceutical effective amount.

3. pharmaceutical composition as claimed in claim 2, it is characterized in that, described pharmaceutical composition comprises the second therapeutical agent further, and described second therapeutical agent is selected from HCV antiviral agent, immunomodulator or anti-infective medicament.

4. pharmaceutical composition as claimed in claim 3, it is characterized in that, described HCV antiviral agent is selected from HCV protease inhibitor or HCV NS5B AG14361.

5. pharmaceutical composition as claimed in claim 2, it is characterized in that, the form of described pharmaceutical composition is aqueous dispersant, liquid, syrup, medicine slurry, suspension, aerosol, control-released agent, quick-dissolving agent, effervescent, freeze-dried, tablet, powder, pill, capsule, delayed release agent or multiparticulates agent.

6. compound as claimed in claim 1 or its pharmacy acceptable salt are preparing the purposes in medicine, and this medicine is used for prevention or treatment HCV infection in the main body needed.

7. purposes as claimed in claim 6, it is characterized in that, wherein said medicine comprises the second therapeutical agent that at least one is selected from HCV antiviral agent, immunomodulator or anti-infective medicament further.

8. purposes as claimed in claim 7, it is characterized in that, wherein HCV antiviral agent is selected from HCV protease inhibitor or HCV NS5B AG14361.